• Fisheries and Aquatic Sciences
• /
• v.19 no.4
• /
• pp.21.1-21.11
• /
• 2016

Background: The evaluation of suitable reference genes as normalization controls is a prerequisite requirement for launching quantitative reverse transcription-PCR (RT-qPCR)-based expression study. In order to select the stable reference genes in abalone Haliotis discus hannai tissues (gill and hepatopancreas) under heavy metal exposure conditions (Cu, Zn, and Cd), 12 potential candidate housekeeping genes were subjected to expression stability based on the comprehensive ranking while integrating four different statistical algorithms (geNorm, NormFinder, BestKeeper, and ${\Delta}CT$ method). Results: Expression stability in the gill subset was determined as RPL7 > RPL8 > ACTB > RPL3 > PPIB > RPL7A > EF1A > RPL4 > GAPDH > RPL5 > UBE2 > B-TU. On the other hand, the ranking in the subset for hepatopancreas was RPL7 > RPL3 > RPL8 > ACTB > RPL4 > EF1A > RPL5 > RPL7A > B-TU > UBE2 > PPIB > GAPDH. The pairwise variation assessed by the geNorm program indicates that two reference genes could be sufficient for accurate normalization in both gill and hepatopancreas subsets. Overall, both gill and hepatopancreas subsets recommended ribosomal protein genes (particularly RPL7) as stable references, whereas traditional housekeepers such as ${\beta}-tubulin$ (B-TU) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were ranked as unstable genes. The validation of reference gene selection was confirmed with the quantitative assay of MT transcripts. Conclusions: The present analysis showed the importance of validating reference genes with multiple algorithmic approaches to select genes that are truly stable. Our results indicate that expression stability of a given reference gene could not always have consensus across tissue types. The data from this study could be a good guide for the future design of RT-qPCR studies with respect to metal regulation/detoxification and other related physiologies in this abalone species.

• Entomological Research
• /
• v.48 no.5
• /
• pp.390-399
• /
• 2018

In order to precisely assess gene expression levels, the suitable internal reference genes must be served to quantify real-time reverse transcription polymerase chain reaction (RT-qPCR) data. For armyworm, Mythimna separata, which reference genes are suitable for assessing the level of transcriptional expression of target genes have yet to be explored. In this study, eight common reference genes, including ${\beta}$-actin (${\beta}$-ACT), 18 s ribosomal (18S), 28S ribosomal (28S), glyceraldehyde-3-phosphate (GAPDH), elongation fator-alpha ($EF1{\alpha}$), TATA box binding protein (TBP), ribosomal protein L7 (RPL7), and alpha-tubulin (${\alpha}$-TUB) that in different developmental stages, tissues and insecticide treatments of M. separata were evaluated. To further explore whether these genes were suitable to serve as endogenous controls, three software-based approaches (geNorm, BestKeeper, and NormFinder), the delta Ct method, and one web-based comprehensive tool (RefFinder) were employed to analyze and rank the tested genes. The optimal number of reference genes was determined using the geNorm program, and the suitability of particular reference genes was empirically validated according to normalized HSP70, and MsepCYP321A10 gene expression data. We found that the most suitable reference genes for the different experimental conditions. For developmental stages, 28S/RPL7 were the optimal reference genes, both $RPL7/EF1{\alpha}$ were suitable for experiments of different tissues, whereas for insecticide treatments, $28S/{\alpha}-TUB$ were suitable for normalizations of expression data. In addition, $28S/{\alpha}-TUB$ were the suitable reference genes because they have the most stable expression among different developmental stages, tissues and insecticide treatments. Our work is the first report on reference gene selection in M. separata, and might serve as a precedent for future gene expression studies.

• KSII Transactions on Internet and Information Systems (TIIS)
• /
• v.8 no.7
• /
• pp.2420-2433
• /
• 2014

Fusing the scores of multibiometrics is a very promising approach to improve the overall system's accuracy and the verification performance. In recent years, there are several approaches towards studying score level fusion of several biometric systems. However, most of them does not consider the genuine and imposter score distributions and result in a higher equal error rate usually. In this paper, a novel score level fusion approach of different biometric systems (dual iris, thermal and visible face traits) based on $Acz{\acute{e}}l$-Alsina triangular norm is proposed. It achieves higher identification performance as well as acquires a closer genuine distance and larger imposter distance. The experimental tests are conducted on a virtual multibiometrics database, which merges the challenging CASIA-Iris-Thousand database with noisy samples and the NVIE face database with visible and thermal face images. The rigorous results suggest that significant performance improvement can be achieved after the implementation of multibiometrics. The comparative experiments also ascertain that the proposed fusion approach outperforms the state-of-art verification performance.

• Nuclear Engineering and Technology
• /
• v.55 no.5
• /
• pp.1757-1762
• /
• 2023

Radiological hazards from external exposure of naturally occurring radioactive materials (NORM) scales residues, generated during the extraction process of oil and gas production in southern Algeria, are evaluated. The activity concentrations of ²²⁶Ra, ²³²Th, and ⁴⁰K were measured using high-purity gamma-ray spectrometry (GeHP). Mean activity concentration of ²²⁶Ra, ²³²Th and ⁴⁰K, found in scale samples are 4082 ± 41, 1060 ± 38 and 568 ± 36 Bq kg^-1, respectively. Radiological hazard parameters, such as radium equivalent (Ra_eq), external and internal hazard indices (H_ex, H_in), and gamma index (I_γ) are also evaluated. All hazard parameter values were greater than the permissible and recommended limits and the average annual effective dose value exceeded the dose constraint (0.3 mSv y^-1). However, for occasionally exposed workers, the dose rate of 0.65 ± 0.02 mSv y^-1 is lower than recommended limit of 1 mSv y^-1 for public.

• Asian-Australasian Journal of Animal Sciences
• /
• v.27 no.4
• /
• pp.471-478
• /
• 2014

Investigating gene expression of immune cells of whole blood or peripheral blood mononuclear cells (PBMC) under polyinosinic:polycytidylic acid (poly I:C) stimulation is valuable for understanding the immune response of organism to RNA viruses. Quantitative real-time PCR (qRT-PCR) is a standard method for quantification of gene expression studies. However, the reliability of qRT-PCR data critically depends on proper selection of reference genes. In the study, using two different analysis programs, geNorm and NormFinder, we systematically evaluated the gene expression stability of six candidate reference genes (GAPDH, ACTB, B2M, RPL4, TBP, and PPIA) in samples of whole blood and PBMC with or without poly I:C stimulation. Generally, the six candidate genes performed a similar trend of expression stability in the samples of whole blood and PBMC, but more stably expressed in whole blood than in PBMC. geNorm ranked B2M and PPIA as the best combination for gene expression normalization, while according to NormFinder, TBP was ranked as the most stable reference gene, followed by B2M and PPIA. Comprehensively considering the results from the two programs, we recommended using the geometric mean of the three genes, TBP, PPIA and B2M, to normalize the gene expression of whole blood and PBMC with poly I:C stimulation. Our study is the first detailed survey of the gene expression stability in whole blood and PBMC with or without poly I:C stimulation and should be helpful for investigating the molecular mechanism involved in porcine whole blood and PBMC in response to poly I:C stimulation.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.21 no.8
• /
• pp.417-428
• /
• 2020

Housekeeping genes are expressed in cells of all organisms and perform basic cellular functions such as energy generation, substance synthesis, cell death, and cell defense. Accordingly, the expression levels of housekeeping genes are relatively constant, and thus they are used as reference genes in gene expression studies, such as protein expression and mRNA expression analysis of target genes. However, the levels of expression of these genes may be different among various tissues or cells and may change under certain circumstances. Therefore, it is important to select the best reference gene for specific gene expression research by exploring the stability of housekeeping gene expression. This review summarizes housekeeping genes found in humans, chickens, pigs, and rats in the literature and estimates expression stability using geNorm, NormFinder, and BestKeeper software. The most suitable reference housekeeping gene can selected based on expression stability according to the experimental conditions of the gene expression study and can thus be applied to data normalization.

• The Plant Pathology Journal
• /
• v.27 no.4
• /
• pp.301-309
• /
• 2011

The filamentous fungus Fusarium graminearum is an important cereal pathogen. Although quantitative realtime PCR (qRT-PCR) is commonly used to analyze the expression of important fungal genes, no detailed validation of reference genes for the normalization of qRT-PCR data has been performed in this fungus. Here, we evaluated 15 candidate genes as references, including those previously described as housekeeping genes and those selected from the whole transcriptome sequencing data. By a combination of three statistical algorithms (BestKeeper, geNorm, and NormFinder), the variation in the expression of these genes was assessed under different culture conditions that favored mycelial growth, sexual development, and trichothecene mycotoxin production. When favoring mycelial growth, GzFLO and GzUBH expression were most stable in complete medium. Both EF1A and GzRPS16 expression were relatively stable under all conditions on carrot agar, including mycelial growth and the subsequent perithecial induction stage. These two genes were also most stable during trichothecene production. For the combined data set, GzUBH and EF1A were selected as the most stable. Thus, these genes are suitable reference genes for accurate normalization of qRT-PCR data for gene expression analyses of F. graminearum and other related fungi.

• The Plant Pathology Journal
• /
• v.35 no.1
• /
• pp.11-18
• /
• 2019

Gaeumannomyces graminis var. tritici is a soil borne pathogenic fungus associated with wheat roots. The accurate quantification of gene expression during the process of infection might be helpful to understand the pathogenic molecular mechanism. However, this method requires suitable reference genes for transcript normalization. In this study, nine candidate reference genes were chosen, and the specificity of the primers were investigated by melting curves of PCR products. The expression stability of these nine candidates was determined with three programs-geNorm, Norm Finder, and Best Keeper. $TUB{\beta}$ was identified as the most stable reference gene. Furthermore, the exopolygalacturonase gene (ExoPG) was selected to verify the reliability of $TUB{\beta}$ expression. The expression profile of ExoPG assessed using $TUB{\beta}$ agreed with the results of digital gene expression analysis by RNA-Seq. This study is the first systematic exploration of the optimal reference genes in the infection process of Gaeumannomyces graminis var. tritici.

• The Pure and Applied Mathematics
• /
• v.15 no.4
• /
• pp.407-414
• /
• 2008

In this paper, we study the self-adjoint second order boundary value problem with integral boundary conditions: (p(t)x'(t))'+f(t,x(t))=0, t $${\in}$$ (0,1), x'(0)=0, x(1) = $${\int}_0^1$$ x(s)g(s)ds. A new result on the existence of positive solutions is obtained. The interesting points are: the first, we employ a new tool-the recent Leggett-Williams norm-type theorem for coincidences; the second, the boundary value problem is involved in integral condition; the third, the solutions obtained are positive.

• The Pure and Applied Mathematics
• /
• v.16 no.2
• /
• pp.213-225
• /
• 2009

This paper deals with the existence of positive solutions for a kind of multi-point nonlinear fractional differential boundary value problem at resonance. Our main approach is different from the ones existed and our main ingredient is the Leggett-Williams norm-type theorem for coincidences due to O'Regan and Zima. The most interesting point is the acquisition of positive solutions for fractional differential boundary value problem at resonance. And an example is constructed to show that our result here is valid.